L9: Generation of Diversity Flashcards Preview

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Flashcards in L9: Generation of Diversity Deck (27)
1

What is the T cell receptor made up of?

2 chains, an alpha chain and a beta chain

2

What is the B cell receptor made up of?

A heavy chain and a light chain
There are 2 light chain genes: kappa and delta (one or the other is included in the BCR expressed by an individual B cell)

3

What processes generate diversity in both B cells and T cells?

1. Multiple copies of each gene segment that makes up the variable joined in different combinations (combinatorial)

2. Addition and subtraction of nucleotides by the recombination process (junctional diversity)

3. Many different combinations of H and L chain V regions (or alpha and beta V regions) that pair to form the antigen binding site (combinatorial)

4

What additional processes add to BCR diversity?

4. Somatic hypermutation occurring in the germinal center

5

How many antigen binding sites do TCRs and BCRs have? What are they made up of?

BCRs: 2 identical antigen binding sites; formed by the heavy chain and the light chain

TCRs: Singular antigen binding site; formed by the alpha and beta chains

6

Where are BCRs expressed?

On surface of B cell or secreted as antibody

7

Why is generation of diversity important?

Survival is dependent upon a wide range of antigen specific receptors, differing in their amino acids sequences at the antigen binding site

But encoding each receptor chain variant in the genome would require more genes than are present in the entire genome

8

What undergoes splicing in TCR and BCR genes?

Both the DNA itself as well as the primary RNA transcript

9

Describe Ig light chain gene construction

Light chain variable region (V_L) is constructed from 2 segments, a variable (V) and joining (J) gene segment; there is a leader peptide (L) that precedes the V gene segment

V and J gene segments in the genomic DNA are joined to form a complete V_L region exon → Light-chain C region (C_L) is encoded in a separate exon and is joined to the V_L exon by splicing of the light-chain RNA to remove the L-to-V and J-to-C introns

10

Describe Ig heavy chain gene construction

Heavy chain V region (V_H) is constructed from 3 gene segments: the diversity (D), V, and J segments

D and J gene segments join → V gene segment joins to the combined DJ sequence → forms a complete V_H exon → During RNA processing of heavy-chain RNA transcript, C-region exons, together w/ leader sequence, are spliced to the V_H domain sequence

11

How many gene segments are there for V, D, and J loci?

Multiple gene segments for each

For instance, for heavy chain there are 45 possible V regions

12

What is intragenic recombination mediated by?

Mediated by the RAG recombinases recognizing RSS (recombination signal sequence)

13

Describe what RAG recombinases do?

RAG recombinase components (RAG-1 and RAG-2) align the segments that will be spliced and excise the intervening DNA including the RSS sequence

Get a signal joint and a coding joint (this gives you VJ region; this is the spliced region that will be expressed)

Rejoining of DNA ends is done by DNA repair enzymes

14

Why does immunodeficiency occur if there is a problem w/ DNA repair enzymes?

Can't complete DNA repair necessary for T and B cell diversity since can't resolve the cuts that the RAG enzymes have made

15

Where are RAG-1 and RAG-2 expressed?

Only in developing lymphocytes
(since don't want it recombining genes in random)

16

Is ligating at the ends of the signal joint and the coding joint precise?

At the signal joint, DNA ligase joins the blunt ends precisely

At the coding join, ligation occurs imprecisely, thus inducing additional variability y into the variable region

17

TdT

(Terminal deoxytransferase)
Adds N (nontemplated) nucleotides at the coding joints; this is a random process

18

What can aberrant rearrangements lead to?

Chromosomal translocations and malignancy

The unique VDJ rearrangement aids in recognizing the presence of a lymphoid malignancy

19

In B cell development, does H chain or L chain gene rearrangement occur first?

VDH rearrangement of heavy chain occurs first

VJ rearragement of light chain then occurs

20

How do nonproductive rearrangements occur?

Added nucleotides at the junctions often disrupt the reading frame, causing a nonproductive rearrangement in 2/3

21

What happens when nonproductive rearrangements occur?

B cell progenitors that do not produce functional immunoglobulin never mature

22

If rearrangement in unsuccessful, what occurs?

Try again since there are 2 alleles

If get it wrong twice, there is cell death

Once able to express heavy chain protein, the rearrangement for light chain protein begins

23

What do all BCR constant regions start off as?

IgM

IgM and IgD seem to be expressed in early B cells

24

What is expressed on the surface of naive B cells?

IgM and IgD

These cell have not yet undergone class-switching

25

In germline TCR genes, are there D regions?

Alpha-chain locus has V and J

Beta-chain locus has V, D, and J

26

If you have defects in VDJ recombination in either RAG or in DNA repair enzymes, what cells are going to have defects?

Both B and T cells

27

Describe TCR rearrangement compared to BCR rearragement

Occurs by the same mechanisms

RAG-1/2 bind to RSS sequences and mediate DNA breakages

Rejoining is mediated by the same DNA repair enzymes

Addition of P and N nucleotides at joints occurs in the same way

Defects in VDJ recombination affect both T and B cells

Somatic hypermutation does not occur after TCR rearrangement